Infectious
pancreatic necrosis virus (IPNV) is an important aquatic pathogen causing a
highly devastating disease in salmonids. The virus has been isolated from over
50 species across the world. For combating the disease, vaccines have been
developed by different recombinant DNA technologies. Production of live virus
vaccines with defined attenuations requires reverse genetics system and
minigenome synthesis to study the attenuation and virus production in vitro systems. Towards this
objective, the two open reading frames of the IPNV (West
Buxton strain) were genetically engineered to replace them with
bacterial chloramphenicol acetyl transferase (CAT) reporter gene while retaining
the non-coding regions (NCR). The minigenome of IPNV without the coding regions
was generated using a modified pUC 19 plasmid and was checked for the
nucleotide correctness by dideoxy chain termination method. Expression of the
reporter gene was verified after transfection studies in susceptible cell line.
The synthesised minigenome is useful in carrying out a number of studies in the
reverse genetics of IPNV.